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Åke Lernmark

Principal investigator

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Epitope analysis of insulin autoantibodies using recombinant Fab

Author

  • C J Padoa
  • N J Crowther
  • J W Thomas
  • T R Hall
  • L M Bekris
  • Carina Törn
  • Mona Landin-Olsson
  • E Ortqvist
  • J P Palmer
  • Åke Lernmark
  • C S Hampe

Summary, in English

Autoantibodies to insulin are often the first autoantibodies detected in young children with type 1 diabetes and can be present before the onset of clinical diabetes. These autoantibodies and their epitopes are, however, not well characterized. We explored the use of monoclonal antibodies and their recombinant Fab as reagents for epitope analysis. In this study we cloned and characterized the recombinant Fab of the insulin-specific monoclonal antibody CG7C7. We found the epitope of this antibody to be located predominantly at the A-chain loop of the insulin molecule. The recombinant Fab was then used to compete for insulin binding against insulin autoantibodies present in sera from patients with type 1 or type 1.5 diabetes. In competition experiments with sera positive for autoantibodies to insulin the recombinant Fab significantly reduced the binding to [I-125]-insulin by sera of type 1 (n = 35) and type 1.5 diabetes [latent autoimmune diabetes in adults (LADA)] (n = 14) patients (P < 0.0001). We conclude that competition between insulin-specific monoclonal antibodies or their recombinant Fab and insulin autoantibodies should prove useful in the epitope analysis of autoantibodies to insulin.

Department/s

  • Diabetes and Celiac Unit
  • Medicine, Lund

Publishing year

2005

Language

English

Pages

564-571

Publication/Series

Clinical and Experimental Immunology

Volume

140

Issue

3

Document type

Journal article

Publisher

British Society for Immunology

Topic

  • Immunology in the medical area

Keywords

  • type 1 diabetes
  • epitopes
  • insulin autoantibody
  • radioligand binding
  • assay
  • recombinant Fab

Status

Published

Research group

  • Diabetes and Celiac Unit

ISBN/ISSN/Other

  • ISSN: 0009-9104